The present invention relates to instruments for chemical reaction control and detection of participating reactants and resultant products, particularly to integrated microfabricated instruments for performing microscale chemical reactions involving precise control of parameters of the reactions, and more particularly to silicon-based and non-silicon based sleeve devices as reaction chambers for chemical reactions and which can be utilized in large arrays of individual chambers for a high-throughput microreaction unit.
Current instruments for performing chemical synthesis through thermal control and cycling are generally very large (table-top) and inefficient, and often they work by heating and cooling of a large thermal mass (e.g., an aluminum block). In recent years efforts have been directed to miniaturization of these instruments by designing and constructing reaction chambers out of silicon and silicon-based materials (e.g., silicon, nitride, polycrystalline silicon) that have integrated heaters and cooling via convection through the silicon.
Microfabrication technologies are now well known and include sputtering, electrode position, low-pressure vapor deposition, photolithography, and etching. These and similar processes can be applied to the fabrication of reaction chambers and their control elements such as heaters, thermocouples, detectors, sensors, electrodes, and other devices that can be used to sense and control the reaction parameters. Examples include magnetic films, thermoelectric films, and electroactive films for reagent manipulation. Additional fabrication techniques include evaporation, extrusion, casting, sintering, injection, forming, pulling, laminating, etc. can be used to microfabricate reaction chambers out of a variety of appropriate materials. Microfabricated devices are formed, for example, on crystalline substrates, such as silicon and gallium arsenide, but may be formed on non-crystalline materials, such as glass, ceramic, metals, or certain polymers. The shapes of crystalline silicon devices, for example, can be precisely controlled since etched surfaces are generally crystal planes, and crystalline materials may be bonded by processes such as fusion at elevated temperatures, anodic bonding, or field-assisted methods.
Monolithic microfabrication technology now enables the production of electrical, mechanical, electromechanical, optical, chemical and thermal devices, including pumps, valves, heaters, mixers, and detectors for microliter to nanoliter quantities of gases, liquids, and solids. Also, optical waveguide probes and ultrasonic flexural-wave sensors can now be produced on a microscale. The integration of these microfabricated devices into a single systems allows for the batch production of microscale reactor-based analytical instruments. Such integrated microinstruments may be applied to biochemical, inorganic, or organic chemical reactions to perform biomedical and environmental diagnostics, as well as biotechnological processing and detection.
The operation of such integrated microinstruments is easily automated, and since the analysis can be performed in situ, contamination is very low. Because of the inherently small sizes of such devices, the heating and cooling can be extremely rapid. These devices have very low power requirement and can be powered by batteries or by electromagnetic, capacitive, inductive or optical coupling.
The small volumes and high surface-area to volume ratios of microfabricated reaction instruments provide a high level of control of the parameters of a reaction. Heaters may produce temperature cycling or ramping; while sonochemical and sonophysical changes in conformational structures may be produced by ultrasound transducers; and polymerizations may be generated by incident optical radiation.
Synthesis reactions, and especially synthesis chain reactions such as the polymerase chain reaction (PCR), are particularly well-suited for microfabrication reaction instruments. PCR can selectively amplify a single molecule of DNA (or RNA) of an organism by a factor of 106 to 109. This well-established procedure requires the repetition of heating (denaturing) and cooling (annealing) cycles in the presence of an original DNA target molecule, specific DNA primers, deoxynucleotide triphosphates, and DNA polymerase enzymes and cofactors. Each cycle produces a doubling of the target DNA sequence, leading to an exponential accumulation of the target sequence.
The PCR procedure involves: 1) processing of the sample to release target DNA molecules into a crude extract; 2) addition of an aqueous solution containing enzymes, buffers deoxyribonucleotide triphosphates (dNTPS), and aligonudeotide primers; 3) thermal cycling of the reaction mixture between two or three temperatures (e.g., 90-96, 72, and 37-55xc2x0 C.); and 4) detection of amplified DNA. Intermediate steps, such as purification of the reaction products and the incorporation of surface-bending primers, for example, may be incorporated in the PCR procedure.
A problem with standard PCR laboratory techniques is that the PCR reactions may be contaminated or inhibited by the introduction of a single contaminant molecule of extraneous DNA, such as those from previous experiments, or other contaminants, during transfers of reagents from one vessel to another. Also, PCR reaction volumes used in standard laboratory techniques are typically on the order of 50 microliters. A thermal cycle typically consists of four stages: heating a sample to a first temperature, maintaining the sample at the first temperature, cooling the sample to a second lower temperature, and maintaining the temperature at that lower temperature. Typically, each of these four stages of a thermal cycle requires about one minute, and thus to complete forty cycles, for example, is about three hours. Thus, due to the large volume typically used in standard laboratory procedures, the time involved, as well as the contamination possibilities during transfers of reagents from one vessel to another, there is clearly a need for microinstruments capable of carrying out the PCR procedure.
Recently, the cycling time for performing the PCR reaction has been reduced by performing the PCR reaction in capillary tubes and using a forced air heater to heat the tubes. Also, an integrated microfabricated reactor has been recently developed for in situ chemical reactions, which is especially advantageous for biochemical reactions which require high-precision thermal cyding, particularly DNA-based manipulations such as PCR, since the small dimensions of microinstrumentation promote rapid cycling times. This microfabricated reactor is described and claimed in copending U.S. application Ser. No. 07/938,106, filed Aug. 31, 1992 now U.S. Pat. No. 5,639,423, entitled xe2x80x9cMicrofabricated Reactorxe2x80x9d, now Pat. No. 5,639,423 issued Jun. 17, 1997, assigned to the same assignee. Also, an optically heated and optically interrograted micro-reaction chamber, which can be utilized, for example, in the integrated microfabricated reactor of the above-referenced copending application Ser. No. 07/938,106, now Pat. No. 5,639,423 has been developed for use in chemical reactors, and is described and claimed in copending U.S. application Ser. No. 08/489,819, filed Jun. 13, 1995, entitled Diode Laser Heated Micro-Reaction Chamber With Sample Detection Meansxe2x80x9d, assigned to the same assignee.
The present invention is directed to a particular geometry of silicon-based and non-silicon-based micro-reactors that have shown to be very efficient in terms of power and temperature uniformity. The micro-reactor of this invention, which is broadly considered as a silicon-based or non-silicon-based sleeve device for chemical reactions, can be effectively utilized in either of the reactor systems of the above-referenced copending applications. The present invention utilizes, for example, doped polysilicon for heating and bulk silicon for convective cooling. The present invention allows the multi-parameter, simultaneous changing of detection window size, in situ detection, reaction volumes, thermal uniformity, and heating and cooling rates. In addition, it enables the use of large arrays of the individual reaction chambers for a high-throughput microreaction unit.
It is an object of the present invention to provide an improved chemical reaction chamber.
A further object of the invention is to provide a silicon-based or non-silicon-based sleeve type chemical reactor.
A further object of the invention is to provide a microfabricated reactor that uses a combination of materials.
A further object of the invention is to provide a chemical reaction chamber that combines to use of doped polysilicon and bulk silicon.
A further object of the invention is to provide a microfabricated chemical reactor having a sleeve reaction chamber with a slot therein for introducing reaction fluids, either directly or via a tube.
A further object of the invention is to provide silicon or non-silicon reaction sleeves that combines critical ratios of materials to provide the proper thermal characteristics.
A further object of the invention is to provide silicon or non-silicon reaction sleeves that combine a critical ratio of materials to provide control of reagents and products.
A further object of the invention is to provide silicon or non-silicon reactions sleeves that combine a critical ratio of materials to provide the proper thermal response.
A further object of the invention is to provide chemical reaction chambers that combines, for example, the use of doped polysilicon and bulk silicon to provide flexibility in thermal and optical properties allowing the implementation into small and large instruments.
Another object of the invention is to provide an interconnected series or array of silicon or non-silicon reaction sleeves thereby providing a flow-through reaction system.
Another object of the invention is to provide a silicon-based reaction sleeve that combines a critical ratio of silicon and silicon nitride to the volume of material to be heated (e.g., liquid) in order to provide uniform heating, yet low power requirement.
Another object of the invention is to provide a sleeve reaction chamber that will allow the introduction of an insert (e.g., plastic) into the reaction sleeve that contains the reaction mixture, thereby elevating any potential materials incompatibility issues.
Another object of the invention is to provide an interconnected series or array of silicon or non-silicon reaction sleeves connected by tubing of polymers, metals, glasses, and ceramics, similar to a xe2x80x9cstring of beadsxe2x80x9d in appearance.
Another object of the invention is to provide an array of individual reaction chambers for a high-throughput microreaction unit.
Another object of the invention is to provide a hand-held instrument that uses sleeve-type reaction chambers with integrated heaters.
Another object of the invention is to provide a reaction chamber with automated detection and feedback control.
Another object of the invention is to provide for artificial intelligence control of reactions in a reaction chamber.
Another object of the invention is to provide pulse-width modulation as a feedback control for reaction chamber.
Another object of the invention is to provide reaction control with magnetic films, thermoelectric films, or electroactive films, such as electrodes.
Another object of the invention is to provide a combination of detection modules, such as electrochemiluminescence, optical, electrical and capacitive.
Another object of the invention is to provide a combined reaction chamber with microelectrophoresis channels.
Another object of the invention is to provide a reaction chamber made from silicon and non-silicon directly coupled with microelectrophoresis channel made of silicon or non-silicon.
Another object of the invention is to provide a sleeve-type reaction chamber with microelectrophoresis channel coupled via an internal liner.
Another object of the invention is to provide a microelectrophoresis detection system based on optical, electrical, or magnetic devices.
Another object of the invention is to provide a microelectrophoresis detection system based on light emitting diodes and photodiodes.
Other objects and advantages of the present invention will become apparent from the following description and the accompanying drawings. Basically, the invention is a silicon-based or non-silicon-based sleeve for chemical reactions. The invention encompasses a chemical reaction chamber that combines, for example, the use of polysilicon for heating and bulk silicon for convective cooling. The reaction chamber may combine a critical ratio of non-silicon and silicon based materials to provide the thermal properties desired. The silicon-based reaction sleeve, for example, may combine a critical ratio of silicon and silicon nitride to the volume of material to be heated in order to provide uniform heating, yet low power requirements. The reaction sleeve of this invention also allows for the introduction therein of a secondary tube or insert that contains the reaction mixture thereby elevating any potential materially incompatibility issues. The present invention is an extension of the above-referenced integrated microfabricated reactor of above-referenced copending application Ser. No. 07/938,106 and the above-referenced optically integrated micro-reaction chamber of above-referenced copending application Ser. No. 08/489,819. The sleeve reaction chamber can be utilized in chemical reaction systems for synthesis or processing of organic, inorganic, or biochemical reactions, such as the polymerase chain reaction (PCR) and/or other DNA reactions (such as the ligose chain reaction), or other synthetic, thermal-cycling-based reactions.